Fig 2: Overexpression of RSK2 attenuates the therapeutic effect of Idebenone on VD rats via the IκBα/NF-κB axis. (A) The learning ability of rats was detected by escape latency test. (B) The spatial memory ability of rats was detected by platform crossing experiment. (C) The changes of swimming speed. (D) the protein expression of RSK2, IκBα, P-IκBα, p65 and p-p65 in the hippocampus of rats detected by WB. (E) The levels of oxidative stress- and inflammation-related factors in hippocampus tissue measured by ELISA. (F) the neuronal damage in the hippocampal cortex evaluated by HE staining. (G) the neuronal damage in CA1 area of hippocampus examined by Nissl staining. N = 10. All experiments were repeated three times independently. The results were shown as mean ± SEM or representative pictures. Unpaired t-test was used for comparison in panels B/E/G, and two-way ANOVA was used for comparison in panels A/C/D. * p < 0.05, vs the Lv-NC.

Fig 3: miR-216a targets RSK2. (A) The potential target genes of miR-216a predicted in several bioinformatics websites. (B) The intersection of potential target genes. (C) The expression of potential target genes in rat hippocampus examined by RT-qPCR. (D) Detection of RSK2 expression in VD rats by RT-qPCR. (E) pmirGLO plasmid mapping and sequences for RSK2-WT and RSK2-MUT. (F) The luciferase activity of WT-RSK2 and MT-RSK2 tested by dual-luciferase assay. All the experiments were repeated three times independently, and the results were averaged. Unpaired t-test was used for comparison between two groups in panel D. Two-way ANOVA was used for comparison in panels C/F. &p < 0.05, vs sham-operated rats; *p < 0.05, vs the Lv-NC; #p < 0.05, vs the NC mimic; &p < 0.05, vs sham-operated rats.

Fig 4: Overexpression of RSK2 attenuates the therapeutic effect of Idebenone on VD rats via the IκBα/NF-κB axis. (A) The learning ability of rats was detected by escape latency test. (B) The spatial memory ability of rats was detected by platform crossing experiment. (C) The changes of swimming speed. (D) the protein expression of RSK2, IκBα, P-IκBα, p65 and p-p65 in the hippocampus of rats detected by WB. (E) The levels of oxidative stress- and inflammation-related factors in hippocampus tissue measured by ELISA. (F) the neuronal damage in the hippocampal cortex evaluated by HE staining. (G) the neuronal damage in CA1 area of hippocampus examined by Nissl staining. N = 10. All experiments were repeated three times independently. The results were shown as mean ± SEM or representative pictures. Unpaired t-test was used for comparison in panels B/E/G, and two-way ANOVA was used for comparison in panels A/C/D. * p < 0.05, vs the Lv-NC.